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Biomedical subjects

I Valverde

Publications and source records attributed to I Valverde.

At least 73 records · Page 4Linked to original sources
PubMed1994

Insulinotropic action of glutamic acid dimethyl ester.

Glutamic acid dimethyl ester (GME; 3.0-10.0 mM) enhanced insulin release evoked by 6.0-8.3 mM D-glucose, 1.0-10.0 mM L-leucine, or 5.0-10.0 mM 2-amino-bicyclo(2,2,1)heptane-2-carboxylic acid, causing a shift to the left of the sigmoidal relationship between insulin output and D-glucose concentration. In the absence of D-glucose, GME also unmasked the insulinotropic potential of glibenclamide. In islets exposed to L-leucine, the insulinotropic action of GME coincided with an early fall and later increase in 86Rb outflow and augmentation of 45Ca outflow from prelabeled islets. The measurement of O2 uptake, NH4+ output, production of 14CO2 from islets prelabeled with [U-14C]palmitate, generation of 14C-labeled amino acids and 14CO2 from the dimethyl ester of either L-[1-14C]glutamic acid or L-[U-14C]glutamic acid, and D-[2-14C]glucose as well as D-[6-14C]glucose oxidation in the presence or absence of GME indicated that the latter ester was efficiently converted to L-glutamate and its further metabolites. The overall gain in O2 uptake represented the balance between GME oxidation and its sparing action on the catabolism of endogenous fatty acids and exogenous D-glucose. It is proposed that GME might represent a new tool to bypass beta-cell defects in D-glucose transport, phosphorylation, and further metabolism and, hence, to stimulate insulin release in experiments conducted in animal models of non-insulin-dependent diabetes mellitus.

Animals↗
PubMed1993

Renal catabolism of truncated glucagon-like peptide 1.

We have searched for the contribution of the kidney to the catabolism of glucagon-like peptide-1 (7-36)amide or tGLP-1 by analyzing the disappearance of the [125I]tGLP-1 both in vivo, from the plasma of bilaterally nephrectomized (BNX), ureteral-ligated (BUL) and normal rats and in vitro from the perfusate of an isolated rat kidney system. Also, we have measured the degradation of the peptide by the isolated renal tubules. Results from in vivo studies demonstrated that the disappearance half-time (t1/2) of [125I]tGLP-1 was significantly lower in the control than in BUL or BNX rats with the metabolic clearance rate (MCR) being higher in the control than in BUL and BNX group; no difference was found for both parameters between BUL and BNX rats. The urinary excretion of the peptide was negligible. The data from the isolated kidney experiments showed a disappearance of the peptide, which was not due to its spontaneous degradation nor to enzymes released from the kidney to the perfusate. Degradation of the peptide also occurred in the presence of isolated tubules. It was dependent upon the concentration of tubules. This could possibly be due to the action of the brush border-associated peptidases. In conclusion, our results demonstrate that, in the rat, the kidney removes the exogenous tGLP-1 from the peripheral circulation, by a mechanism that involves glomerular filtration and tubular catabolism.

Animals↗
PubMed1993

Respiratory, ionic, and functional effects of succinate esters in pancreatic islets.

The methyl esters of succinic acid were introduced a few years ago as new potent insulin secretagogues. In the present study, they were found to increase O2 uptake by rat islets incubated in the absence or presence of D-glucose; to decrease 86Rb outflow from prelabeled islets; to stimulate biosynthetic activity in the islets, with a preferential effect on the synthesis of proinsulin; to inhibit 45Ca efflux from prelabeled islets perifused in the absence of extracellular Ca2+ but to augment 45Ca net uptake and to cause a biphasic stimulation of 45Ca outflow in islets incubated or perifused in the presence of extracellular Ca2+; and to evoke a biphasic stimulation of insulin release. The insulinotropic action of these methyl esters coincided with a shift to the left of the sigmoidal relationship between insulin output and D-glucose concentration, was concentration related in the 2-10 mM range, failed to be duplicated by succinic acid, displayed both Ca2+ dependency and resistance to a lowering of extracellular pH, and was operative in the absence of D-glucose whether or not the islets were stimulated by non-nutrient secretagogues. It is concluded that the respiratory, cationic, biosynthetic, and secretory responses of the islets to succinate methyl esters display the characteristic features usually encountered in the process of nutrient-stimulated insulin release.

Animals↗
PubMed1993

Presence and characterization of glucagon-like peptide-1(7-36) amide receptors in solubilized membranes of rat adipose tissue.

Specific binding of [125I]glucagon-like peptide-1(7-36)amide ([125I]GLP-1(7-36)amide) to solubilized rat adipose tissue membranes was found to be dependent on temperature, time, and membrane protein concentration and readily dissociated. GLP-1(1-36)amide, GLP-2, or glucagon (10(-6) M) did not compete with [125I]GLP-1(7-36)amide binding. Half-maximal binding was achieved with 8 x 10(-10) M unlabeled GLP-1(7-36)amide, and the Scatchard plot revealed the presence of high and low affinity binding sites with Kd values of approximately 0.6 and 20 nM, respectively. The binding capacity of [125I]GLP-1(7-36)amide was about 3 times higher than that of [125I]glucagon, while the high affinity Kd and the half-maximal binding of the two peptides were similar. The presence and abundance of GLP-1(7-36)amide receptors in fat tissue together with the previous findings that the peptide stimulates glycerol and cAMP production in rat adipocytes and stimulates fatty acid synthesis in explants of rat adipose tissue open the possibility that this insulinotropic intestinal peptide may also be involved in the regulation of lipid metabolism in health and disease.

Adipose Tissue↗
PubMed1993

Presence of glucagon and glucagon-like peptide-1-(7-36)amide receptors in solubilized membranes of human adipose tissue.

Specific receptors for glucagon and for glucagon-like peptide-1 (GLP-1) (7-36)amide have been found in solubilized human adipose membranes. The 50% inhibition dose of the corresponding unlabeled peptide was near to their physiological levels [ID50, 0.5 nmol/L for glucagon and 1.0 nmol/L for GLP-1(7-36)amide;]. In both cases, the presence of high affinity receptors was evident [Kd, 0.5 and 0.7 nmol/L for glucagon and GLP-1(7-36)amide, respectively]; the high affinity maximal binding capacity for GLP-1(7-36)amide was higher than that for glucagon (893 and 117 fmol/mg solubilized fat membranes, respectively). Glucagon at 10(-6) mol/L did not compete with the [125I]GLP-1(7-36)amide binding, nor did GLP-1(7-36)amide (10(-6) mol/L) compete with that of [125I]glucagon. The relative abundance of GLP-1(7-36)amide receptors in human adipose tissue is further support for a direct and probably important action of this peptide in the metabolism of the fat cell.

Adipose Tissue↗
PubMed1992

[Trophic activity of neurotensin in massive intestinal resection].

Regulatory peptides are among the main factors affecting the proliferative adaptive process of the small bowel; its mechanisms of action remains unknown. An 80% small bowel resection was done in Wistar rats (250-300 g) randomly assigned to two groups: control and neurotensin (300 mu/kg). Jejunal and ileal samples were taken on day 14 post-resection and histomorphometric (villous length) and proliferative (DNA content) measures were obtained. Results show an increase in villous length in the neurotensin group but no changes in DNA content were observed. We conclude that neurotensin increases intestinal mucosal growth after massive intestinal resection in the rat.

Animals↗
PubMed1991

Metabolic labelling and partial characterization of glycophospholipids in pancreatic islet cells.

Insulin action is thought to be mediated by an inositol-, glucosamine- and galactose-containing oligosaccharide liberated by phosphodiesterase hydrolysis of a glycosyl-phosphatidylinositol. This oligosaccharide inhibits insulin biosynthesis and secretion in pancreatic islets. In the present study, two main glycolipids (peak I and II) were resolved by sequential TLC of lipids extracted from islet cells labelled with tritiated glucosamine, galactose or myristate. The two glycolipids displayed comparable sensitivity to beta-galactosidase but differed from one another by their sensitivity to phosphatidylinositol-specific phospholipase C. Moreover, structural heterogeneity within each peak was suggested by their partial resistance to nitrous acid deamination. These findings support the presence in islet cells of glycolipids similar to those currently considered as a possible postreceptor target for insulin in other cell types.

Animals↗
PubMed1991

Relationship between D-glucose oxidation and glycolysis in tumoral pancreatic islet cells with either rapid or decreased cell growth.

The relationship between glucose oxidation and glycolysis was examined in tumoral pancreatic islet cells with either rapid or decreased cell growth. In the control RINm5F cells, the utilization of D-[5-3H]glucose progressively augmented at increasing concentrations of D-glucose (0.2 to 16.7 mM), whereas D-[6-14C]glucose oxidation reached its maximal value at about 3.0 mM D-glucose. The 14CO2/3HOH ratio progressively decreased at increasing hexose concentrations, being about 5-times higher at 0.2 than 10.0-16.7 mM. These experiments were repeated in cells cultured in the presence of D,L-alpha-difluoromethylornithine, which inhibits RINm5F cell proliferation. At 0.2 mM D-glucose, 14CO2 output was severely decreased but 3HOH production little affected, resulting in a marked fall of the 14CO2/3HOH ratio. On the contrary, at 16.7 mM D-glucose, 3HOH formation was more markedly decreased than 14CO2 output, resulting in an increased 14CO2/3HOH ratio. These findings provide a model for the interference of cell growth with D-glucose metabolism in islet cells. Rapid growth causes a preferential stimulation of the Krebs cycle relative to glycolysis at a low glucose concentration, whilst increasing glycolysis more than oxidation at high concentrations of the hexose.

Animals↗
PubMed1990

Renal catabolism of human glucagon-like peptides 1 and 2.

The renal catabolism of [125I]glucagon-like peptide 1 (GLP-1) and [125I]glucagon-like peptide 2 (GLP-2) has been studied both in vivo, by the disappearance of these peptides from the plasma of bilaterally nephrectomized (BNX), ureteral-ligated (BUL) or normal rats, and in vitro, analyzing their catabolism by the isolated, perfused rat kidney. Results from in vivo studies demonstrated that half-disappearance time for both peptides was lower in controls than in BUL rats, and this value in BUL rats was not significantly different from that in BNX rats. In addition, metabolic clearance rate of GLP-1 was higher in control rats than in the other two groups of animals. Urinary clearance rate of both peptides was negligible. In isolated kidney experiments, values for organ clearance of both [125I]GLP-1 and [125I]GLP-2 were similar to those of inulin clearance, which represents the glomerular filtration rate. Urinary clearance of trichloroacetic acid precipitable radioactivity represented less than 1% of total clearance. In conclusion, these results demonstrate a significant role for the kidney in the plasma removal of [125I]GLP-1 and [125I]GLP-2 by a mechanism that involves glomerular filtration and tubular catabolism.

Animals↗
PubMed1990

Ketone bodies and islet function: 45Ca handling, insulin synthesis, and release.

D-(-)-beta-Hydroxybutyrate and acetoacetate cause a rapid, sustained, and rapidly reversible stimulation of insulin release from rat pancreatic islets incubated in the presence, but not absence, of D-glucose. This coincides with stimulation of both proinsulin biosynthesis and 45Ca net uptake. The ketone bodies also decrease 45Ca outflow from prelabeled islets perifused in the absence of Ca2+ and, in contrast, enhance effluent radioactivity in the presence of Ca2+. In the presence of D-glucose, the secretory response to D-(-)-beta-hydroxybutyrate is concentration related in the 2.5-20 mM range, abolished in the absence of Ca2+ or presence of KCN, and enhanced by theophylline and forskolin. It corresponds grossly to a shift to the left of the sigmoidal curve relating insulin output to the ambient concentration of D-glucose. The secretory, biosynthetic, and cationic response to acetoacetate is less marked than that evoked by an equimolar concentration of D-(-)-beta-hydroxybutyrate. These features are compatible with the view that the insulinotropic action of ketone bodies would be causally linked to their metabolism in islet cells.

3-Hydroxybutyric Acid↗
PubMed1990

Binding of [3H]cytochalasin B to tumoral islet cells.

Tumoral pancreatic islet cells of the RINm5F line are equipped with two classes of [3H]cytochalasin B binding sites with respective Kd of 0.4 and 7 microM. The binding of the fungal metabolite and its dissociation from the binding sites display rapid time courses. The binding is inhibited by D-glucose, more than by L-glucose, by phlorizin and by cytochalasin E. These findings are considered in the light of the dual action of cytochalasin B upon hexose transport and motile activity in islet cells.

Adenoma, Islet Cell↗
PubMed1989

Binding of ouabain to normal and tumoral pancreatic islet cells.

The binding of [3H(G)]ouabain was examined in both rat pancreatic islets and tumoral islet cells (RINm5F line). In the islets, the specific binding was considered too low to allow for extensive investigations, taking into account the limited number of islets readily available. In the RINm5F cells, which bound more ouabain than normal islet cells, the time course for binding, its temperature dependency, its competitive inhibition by K+, the number of binding sites, their affinity, and the turnover of ATP per binding site were all similar to those found in other cell types. An incomplete dissociation of [3H(G)]ouabain by unlabelled glycoside and the stimulation of binding by D-glucose were noticed, both phenomena being possibly attributable, in part at least, to the functional response of the RINm5F cells to D-glucose with resulting activation of exocytosis-endocytosis coupling.

Animals↗
PubMed1989

Stimulus-secretion coupling of arginine-induced insulin release. Metabolism of L-arginine and L-ornithine in pancreatic islets.

Exogenous L-arginine and L-ornithine rapidly accumulate in rat pancreatic islets. L-Arginine is converted to L-ornithine and urea. Endogenous or exogenous L-ornithine generates di- and polyamines, the putrescine turnover being faster than that of spermidine and spermine. However, the major pathway for L-ornithine metabolism consists of its transamination to L-glutamaldehyde and further conversion to L-glutamate. The amines and L-glutamate derived from exogenous L-ornithine are incorporated into islet proteins at the intervention of transglutaminase and cycloheximide-sensitive biosynthetic processes, respectively. These findings suggest the hypothesis that the insulinotropic action of L-arginine and L-ornithine could somehow be related to the metabolism of these cationic amino acids in islet cells.

Animals↗
PubMed1989

Effect of a phospho-oligosaccharidic putative insulin messenger on insulin release in rats.

The phospho-oligosaccharide extracted from rat liver and supposed to act as the insulin second messenger inhibits glucose-stimulated insulin release. In the present study, this phospho-oligosaccharide was found not to affect D-[U-14C]glucose oxidation and 45Ca net uptake, but to inhibit insulin release evoked by either D-glucose or 2-ketoisocaproate in isolated rat islets. The relative extent of the latter inhibition was unaffected by either the concentration of D-glucose or the presence of dibutyryl-cyclic AMP, forskolin or glucagon in the incubation medium. At variance with the inhibitory effect of clonidine, that of the phospho-oligosaccharide was resistant to both blockade of alpha 2-adrenergic receptors or pre-treatment with the toxin of Bordetella pertussis. It is speculated, therefore, that such a phospho-oligosaccharide might interfere with a distal event in the insulin secretory sequence.

Animals↗
PubMed1989

Stimulus-secretion coupling of arginine-induced insulin release: metabolism of L-arginine and L-ornithine in tumoral islet cells.

The metabolism of L-arginine and L-ornithine was examined in tumoral islet cells of the RINm5F line and compared to the situation previously characterized in normal rat islets. The maximal velocity of arginase in cell homogenates, as well as either the production of 14C-urea or the steady-state content of 14C-labelled ornithine in intact cells exposed to L-[U-14C]arginine were about one order of magnitude lower in tumoral than normal islet cells. The activity of ornithine-glutamate transaminase was similar in both cell types, and this coincided with a comparable rate of 14C-labelled L-glutamate generation by intact cells exposed to L-[1-14C]ornithine. Despite a comparable cell content in 14C-labelled ornithine of normal and tumoral cells exposed to exogenous ornithine, the rate of di- and polyamine generation was about one order of magnitude higher in tumoral than normal islet cells, this coinciding with a much higher activity of ornithine decarboxylase in RINm5F cell than islet homogenates.

Adenoma, Islet Cell↗
PubMed1989

Inhibition by a nonmetabolized analogue of L-leucine of protein biosynthesis in tumoral pancreatic islet cells.

The effect of L-leucine, its deaminated metabolite 2-ketoisocaproate and its nonmetabolized analogue b(+/-)2-aminobicyclo[2,2,1]-heptane-2-carboxylic acid (BCH) upon protein labelling was examined in tumoral islet cells (RINm5F line) exposed to L-[4-3H]phenylalanine or L-[3-3H]serine. The interpretation of the results, in terms of changes in biosynthetic activity, was obscured by a possible interference of the tested nutrients with the uptake and further metabolism of the tracer tritiated amino acids. Nevertheless, when the cells were preincubated with the nutrient secretagogues and then incubated in the sole presence of L-[3-3H]serine, BCH, but not L-leucine or 2-ketoisocaproate, still inhibited protein labelling, this coinciding with a decrease in the ratio between TCA-precipitable and total radioactivity in the RINm5F cells. The inhibitory action of BCH was antagonized, to a limited extent, by D-glucose. It is proposed that BCH could be used as a tool to interfere with the function and growth of insulinoma cells.

Adenoma, Islet Cell↗
PubMed1989

Glucagon storage, release and degradation by tumoral islet cells (RINm5F line).

Tumoral islet cells of the RINm5F line contain and release glucagon. Over 90 min incubation, the secreted hormone represents about 50% of the final cell content (1.4 +/- 03 fg/cell). L-arginine augments modestly glucagon release, the relative magnitude of such an effect being inversely related to the concentration of D-glucose. The immunoreactive material secreted by the cells consists of both true glucagon (59-64%) and larger peptides. The RINm5F cells also release a proteolytic factor able to degrade exogenous glucagon or insulin. These results reveal that glucagon producing tumoral cells display several abnormal features in their secretory behaviour.

Adenoma, Islet Cell↗